Purification and High-Resolution Top-Down Mass Spectrometric Characterization of Human Salivary α-Amylase

摘要

1. Method based on amylase and glycogen interaction is the preferred method with the highest purity and reproducibility. 2. We have identified the truncation of the N-terminal 15 amino acids and formation of pyroglutamic acid at the new N-terminus. 3. We have precisely mapped 5 S-S bond positions in HSAMY by top-down MS in combination with partial reduction by TCEP. 4. Our study demonstrates that top-down MS combined with affinity purification and partial reduction is a powerful new method for rapid purification and complete characterization of proteins with complex, overlapping S-S positions.